Surface-induced aggregation of ferritin. Kinetics of adsorption to a hydrophobic surface

Abstract

The adsorption of ferritin from a water solution to a hydrophobic methylised quartz surface was studied by transmission electron microscopy, allowing direct examination of the iron core of the molecule without further preparation. The initial adsorption was seen to result in small clusters of molecules, the number of sites/cm(2) being concentration dependent. The adsorption process continued via cluster growth. The rate of adsorption increased and the process became mass transport limited. The clusters formed initially had low fractal dimensions (D approximately 1.0) and a coordination number, cn of 2.6-2.8, which increased with time. These clusters were abruptly restructured at a coordination number of 3.5, and the apparent rate of adsorption decreased during the reorganisation of the adsorbed layer. Finally, an equilibrium level was reached which was stable for at least 24 h. The distribution of ferritin molecules at equilibrium was in clusters with a fractal dimension of D = 1.14 +/- 0.16 and D= 1.33 +/- 0.08, respectively, for ferritin concentrations in the bulk of 10 and 100 microg/ml. Rinsing of adsorbed ferritin layers with buffered salt solution resulted in a rapid transient condensation of the clusters, but the net dissociation of protein was slow with the rate of dissociation being proportional to the logarithm of time. The condensed clusters were slowly restructured to linear polymers of ferritin molecules with a coordination number of 1.9 after 24 h of rinsing. The dissociation of protein molecules continued slowly for more than 3 days of rinsing. The results of the present study indicate that the rate of protein adsorption and desorption is strongly related to the supramolecular structure of the adsorbed protein film. Dense clusters of protein are not stable and this phenomenon may explain the formation of a dynamic equilibrium in spite of the fact that protein adsorption to a solid phase may appear to be practically irreversible.